2D MEMBRANE PROTEIN CRYSTALLIZATION USING DNA TEMPLATE

使用 DNA 模板进行 2D 膜蛋白结晶

• 批准号: 7721145
• 负责人: Hao Yan
• 金额: $ 1.62万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721145
• 关键词: Biotin; Cellular Membrane; Computer Retrieval of Information on Scientific Projects Database; Cryoelectron Microscopy; Crystallization; Crystallography; DNA; Dimensions; Drug Delivery Systems; Electrostatics; Funding; Goals; Grant; Histidine; Institution; Integral Membrane Protein; Lipids; Membrane; Membrane Proteins; Methods; Molecular Structure; Nickel; Periodicals; Periodicity; Proteins; Research; Research Personnel; Resolution; Resources; Source; Streptavidin; Structure; Technology; United States National Institutes of Health; design; interest; macromolecule; self assembly; two-dimensional

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our goal is to use self-assembled two-dimensional (2D) DNA lattices to direct periodic assembly of membrane proteins to obtain high quality crystals. In recent years, cryo-EM crystallography has been used to obtain high-resolution structures from 2D crystals of protein molecules provided that quality 2D crystals can be achieved. Currently the methods for 2D crystallization of macromolecules heavily rely on unpredictable interaction between the surfaces of the protein molecules (e.g. hydrophobic and electrostatic interaction). Methods have been developed to use biotin-tagged or nickel chelated lipids to organize streptavidin or histidine-tagged proteins into 2D crystal. However, controlled spacing or periodicity of the protein molecule is still hard to control in these designs. Such control is needed to organize proteins of various dimensions. By combining the expertise from Allen on 3D membrane crystallography and Yan on DNA self-assembly, we propose to develop a robust and modular technology to use self-assembled 2D DNA lattices to organize transmembrane proteins into periodical 2D crystals. This modular technology will enable us to rationally organize the protein of interest into an ordered 2D crystal which will facilitate the elucidation of their molecular structures using cryo-electron microscopy studies. Structural studies of membrane proteins will help us understand cellular membrane functions and will have significant impact in designing drug delivery across the membrane.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 我们的目标是使用自组装的二维（2D）DNA晶格直接定期组装膜蛋白来获得高质量的晶体。近年来，只要可以实现质量的2D晶体，冷冻EM晶体学已用于从蛋白质分子的2D晶体中获得高分辨率结构。目前，大分子的2D结晶方法严重依赖于蛋白质分子表面之间的不可预测的相互作用（例如疏水和静电相互作用）。已经开发了使用生物素标签或镍螯合脂质将链霉亲和蛋白标记的蛋白组织到2D晶体中的方法。但是，在这些设计中，仍难以控制蛋白质分子的受控间距或周期性。需要这样的控制来组织各种维度的蛋白质。通过将Allen的专业知识结合到3D膜晶体学上的专业知识和DNA自组装上的YAN，我们建议开发一种可靠的模块化技术，以使用自组装的2D DNA晶格将跨膜蛋白组织到周期性的2D晶体中。这种模块化技术将使我们能够合理地将感兴趣的蛋白质组织成有序的2D晶体，该晶体将促进使用冷冻电子显微镜研究促进其分子结构。膜蛋白的结构研究将有助于我们了解细胞膜功能，并将在整个膜上递送药物方面产生重大影响。